Greater Addition of Neurons to the Olfactory Bulb Than to the Cerebral Cortex of Eulipotyphlans but Not Rodents, Afrotherians Or Primates
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ORIGINAL RESEARCH ARTICLE published: 11 April 2014 NEUROANATOMY doi: 10.3389/fnana.2014.00023 Greater addition of neurons to the olfactory bulb than to the cerebral cortex of eulipotyphlans but not rodents, afrotherians or primates Pedro F. M. Ribeiro 1,2,PaulR.Manger3, Kenneth C. Catania 4, Jon H. Kaas 5 and Suzana Herculano-Houzel 1,2* 1 Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil 2 Instituto Nacional de Neurociência Translacional, Ministério de Ciência e Tecnologia, São Paulo, Brasil 3 Department of Anatomy, University of the Witwatersrand, Johannesburg, South Africa 4 Department of Biology, Vanderbilt University, Nashville, TN, USA 5 Department of Psychology, Vanderbilt University, Nashville, TN, USA Edited by: The olfactory bulb is an evolutionarily old structure that antedates the appearance of a Chet C. Sherwood, George six-layered mammalian cerebral cortex. As such, the neuronal scaling rules that apply to Washington University, USA scaling the mass of the olfactory bulb as a function of its number of neurons might be Reviewed by: shared across mammalian groups, as we have found to be the case for the ensemble Gordon M. Shepherd, Yale University School of Medicine, USA of non-cortical, non-cerebellar brain structures. Alternatively, the neuronal scaling rules Emmanuel Paul Gilissen, Royal that apply to the olfactory bulb might be distinct in those mammals that rely heavily Museum for Central Africa, Belgium on olfaction. The group previously referred to as Insectivora includes small mammals, *Correspondence: some of which are now placed in Afrotheria, a base group in mammalian radiation, Suzana Herculano-Houzel, Instituto and others in Eulipotyphla, a group derived later, at the base of Laurasiatheria. Here de Ciências Biomédicas, Universidade Federal do Rio de we show that the neuronal scaling rules that apply to building the olfactory bulb differ Janeiro, Av. Brigadeiro Trompowski, across eulipotyphlans and other mammals such that eulipotyphlans have more neurons s/n, Ilha do Fundão, Rio de Janeiro – concentrated in an olfactory bulb of similar size than afrotherians, glires and primates. RJ, 21941-590 Brazil Most strikingly, while the cerebral cortex gains neurons at a faster pace than the olfactory e-mail: [email protected] bulb in glires, and afrotherians follow this trend, it is the olfactory bulb that gains neurons at a faster pace than the cerebral cortex in eulipotyphlans, which contradicts the common view that the cerebral cortex is the fastest expanding structure in brain evolution. Our findings emphasize the importance of not using brain structure size as a proxy for numbers of neurons across mammalian orders, and are consistent with the notion that different selective pressures have acted upon the olfactory system of eulipotyphlans, glires and primates, with eulipotyphlans relying more on olfaction for their behavior than glires and primates. Surprisingly, however, the neuronal scaling rules for primates predict that the human olfactory bulb has as many neurons as the larger eulipotyphlan olfactory bulbs, which questions the classification of humans as microsmatic. Keywords: olfactory bulb, cortical expansion, mosaic evolution, olfaction INTRODUCTION in the species examined—but not in Insectivora (Stephan and The olfactory bulb is an evolutionarily ancient structure of the Andy, 1969; Jolicoeur et al., 1984). This seemed consistent both forebrain, found in nearly all vertebrates, including jawless fishes with a basal position of Insectivora and with the progressive view (Butler and Hodos, 2005). The olfactory bulb thus precedes the of evolution still in vogue in the late 20th century. appearance of a well-developed mammalian cerebral cortex, and At that time, Insectivora was a “scrap basket for small mam- its expansion drove the initial increase in relative brain size in mals” (Simpson, 1945),andviewedasanearlyoffshootin mammalian evolution (Rowe et al., 2011). However, the olfac- the placental mammal radiation, based on morphological cri- tory bulb has been shown to scale more slowly than the cerebral teria. In contrast, recent molecular data (Stanhope et al., 1998; cortex in a number of modern mammalian species, although Madsen et al., 2001; Murphy et al., 2001) provided convincing independently of the volume of the cerebral cortex across pri- evidence that the group was polyphyletic, and actually consisted mates (Stephan and Andy, 1969). While the relative volume of the of two orders: afrosoricida (tenrecs and golden moles), under cerebral cortex when regressed onto body mass was once shown to the superorder Afrotheria (Stanhope et al., 1998; Madsen et al., increase with brain volume across living mammalian species, and 2001; Murphy et al., 2001), and Eulipotyphla (hedgehogs, moles, to increase the most amongst primates (leading to the concept of shrews, solenodons; Waddell et al., 1999; Douady et al., 2002; cortical expansion in evolution; Stephan and Andy, 1969), the rel- Waddell and Shelley, 2003). Eulipotyphlans were later grouped ative volume of the olfactory bulb decreases by the same measure into the Laurasiatheria clade, which includes bats, cetartiodactyls, Frontiers in Neuroanatomy www.frontiersin.org April 2014 | Volume 8 | Article 23 | 1 Ribeiro et al. Numbers of neurons in the olfactory bulb carnivores, pangolins, and perissodactyls (Madsen et al., 2001; MATERIALS AND METHODS Murphy et al., 2001). Thus, in contrast from the original view that We applied the isotropic fractionator (Herculano-Houzel and placed insectivores as a whole as an early branch at the root of Lent, 2005) to determine the cellular composition of the olfac- placental mammals, this place is now occupied by Afrotherians tory bulbs of 10 glire species and 6 primate and the closely related (which do include golden moles, some of the original insecti- Scandentia species. Additionally, we compare our data to the cel- vores) and Xenarthra (Madsen et al., 2001; Murphy et al., 2001). lular composition of the olfactory bulbs of 5 insectivore species The remainder of the original insectivores, now known to be (Sarko et al., 2009) and 3 afrotherian species (Neves et al., 2014), eulipotyphlans, are considered a later offshoot, at the base of analyzed previously in the same manner. Laurasiatheria (Madsen et al., 2001; Murphy et al., 2001). We have previously demonstrated that the scaling rules that ANIMALS relate the mass of the cerebral cortex and cerebellum to their A total of 73 olfactory bulbs were collected from primates, rodents numbers of neurons differ across primates, glires and eulipoty- and one scandentia whose brains were prepared for the exami- phlans (reviewed in Herculano-Houzel, 2011). While in primates nation of cerebral structures in unrelated experiments. From pri- these structures scale linearly in mass with their number of mates, 25 olfactory bulbs were collected from 6 galagos (Otolemur neurons (Herculano-Houzel et al., 2007), in glires (rodents and Garnettii, 11 bulbs), 3 marmosets (Callithrix jacchus, 5 bulbs), lagomorphs) they grow in mass with the number of neurons 3owlmonkeys(Aotus trivirgatus, 6 bulbs), one rhesus monkey raised to the power of 1.7 and 1.2, respectively, which indicates (Macaca mulatta, 1 bulb), and one mouse lemur (Microcebus sp., that average neuronal size increases with number of neurons 2 bulbs). From Glires (rodents plus lagomorphs), we collected a (Herculano-Houzel et al., 2006, 2011). In eulipotyphlans, the total of 32 olfactory bulbs from 9 gray squirrels (Sciureus caroli- cerebral cortex scales in mass faster than it gains neurons, as it nensis, 9 bulbs), 5 Wistar rats (Rattus norvegicus, 5 bulbs), 2 agouti does in rodents, while the cerebellum increases in mass as a lin- (Dasyprocta primnolopha, 2 bulbs), 2 swiss mice (Mus musculus, ear function of its numbers of neurons, as in primates (Sarko 4 bulbs), 2 golden hamsters (Mesocrycetus auratus, 2 bulbs), 2 et al., 2009). These differences suggest that the cerebral cortex and guinea pigs (Cavia porcellus, 2 bulbs), 2 capybaras (Hydrochoerus cerebellum of primate, glires and eulipotyphlan brains are built hydrochoeris, 2 bulbs), 1 rabbit (Oryctolagus cuniculus, 1 bulb), according to cellular scaling rules that have diverged from their 3 naked mole-rats (Heterocephalus glaber,3bulbs)andone common ancestor. In contrast, the cellular scaling rules that apply Cayenne spiny rat (Proechimys guyannensis,2bulbs);someof to the remaining areas of the brain, excluding the olfactory bulb the present data on olfactory bulbs were published previously in (that is, the ensemble of brainstem, diencephalon and basal gan- Herculano-Houzel et al. (2011). We also collected 16 olfactory glia), appear to be more similar across these three mammalian bulbsfrom10treeshrews(Tupaia glis). All animals were adults orders, and might possibly be shared among them (Sarko et al., at the time of the experiments. All non-human primates, except 2009; Herculano-Houzel, 2011). This raises the possibility that for the mouse lemur, were raised in a colony in the Department of the olfactory bulb, being a structure as evolutionarily ancient as Psychology at Vanderbilt University. The mouse lemur brain was the brainstem and antedating the expansion of the cerebral cor- acquired from the Duke University Lemur Center. tex, might also vary in mass according to neuronal scaling rules Insectivore data were obtained from Sarko et al. (2009) and that are shared across mammalian orders. consist of the species averages of 34